WO2022037617A1 - Cyanotriazine derivative, preparation method therefor, and application thereof - Google Patents

Abstract

A compound represented by structural formula (I) and a pharmaceutically acceptable salt thereof and a pharmaceutical composition containing them, capable of being used as agonists of a thyroid hormone receptor -β, capable of being used for treating diseases such as obesity, hyperlipidemia, hypercholesterolemia, and diabetes, and also for treating other diseases such as NASH, atherosclerosis, cardiovascular diseases, hypothyroidism, thyroid cancer, and other related disorders and diseases.

Description

Nitrile triazine derivatives and preparation method and application thereof technical field

The invention relates to a thyroid hormone receptor-beta (THR-beta) subtype agonist derivative, a preparation method and its medical application. The derivatives of the present invention or the pharmaceutical combination containing the derivatives of the present invention can be used to treat related diseases. For example, it can be used for obesity, hyperlipidemia, hypercholesterolemia, diabetes, etc.; it can also be used for the treatment of other diseases, including NASH (nonalcoholic steatohepatitis, non-alcoholic fatty liver), hepatic steatosis, atherosclerosis Sclerosis, cardiovascular disease, hypothyroidism, thyroid cancer and related conditions, etc.

Background technique

Thyroid hormones play a key role in normal growth and development as well as maintaining metabolic homeostasis (Physiological Reviews, 2001, 81(3), 1097-1042; Biochimica et Biophysica Acta 1812 (2011) 929-937). Circulating levels of thyroid hormones are tightly regulated by feedback mechanisms in the hypothalamic/pituitary/thyroid axis. Thyroid dysfunction can lead to hypothyroidism or hyperthyroidism. In addition, thyroid hormones have important effects on heart function, body weight, metabolism, metabolic rate, body temperature, cholesterol, bones, muscles, and behavior.

Thyroid hormone is produced by the thyroid gland and secreted into the circulatory system in two distinct forms: 3,5,3',5'-tetra-iodo-L-thyronine (T4) and 3,5,3'-tris -Iodo-L-thyronine (T3). T4 is the main form secreted by the thyroid. T3 is the more physiologically active form of existence. T4 can be converted to T3 by tissue-specific deiodinase. This tissue-specific deiodinase is present in all tissues, but mainly in the liver and kidney. The biological activity of thyroid hormones is mediated by thyroid hormone receptors (THRs) (Endocrine Reviews, 1993, 348-399). THR belongs to a superfamily known as nuclear receptors. The major thyroid receptor subclasses include: alpha1, alpha2, beta1, and beta2. Thyroid hormone receptors α1, β1 and β2 can directly bind to the thyroid hormone T3, while α2 can interact with non-T3 (Advances in Developmental Biology. 2006, 16, 1-31). Studies have shown that thyroid hormone receptor subtypes can differ in their contribution to specific physiological responses. For example, THR-beta1 plays an important role in the regulation of TRH (thyrotropin-releasing hormone) and the action of thyroid hormones in the liver. THR-β2 plays a major role in regulating TH (thyroid stimulating hormone). THR-β1 plays an important role in regulating heart rate. For example, thyroid hormones increase metabolic rate, oxygen consumption, and heat release. thereby reducing weight. Weight loss will have a beneficial effect on obese patients by improving obesity-related comorbidities, and may also have a beneficial effect on glycemic control in obese patients with type 2 diabetes (J. Clin. Invest., 1999, 104:291 -300).

Obviously, by activating THR-β1, the corresponding biological function can be changed, so as to achieve some expected therapeutic purpose. In view of the rising prevalence of obesity and the diseases caused by obesity, including but not limited to diabetes, metabolic syndrome and atherosclerotic vascular disease, there is an urgent need for compounds capable of treating these diseases. Therefore, thyroid hormone analogs with increased thyroid hormone receptor beta-selectivity and/or tissue selection are receiving extensive attention. This THR-beta selective agonist produces modest reductions in body weight, lipids, cholesterol, and certain lipoproteins, with reduced effects on cardiovascular function or normal function of the hypothalamic/pituitary/thyroid axis (J.Med Chem. 2014, 57, 3912-3923; Expert Opin Investig Drugs. 2004, 13(5): 489-500).

Madrigal Pharmaceuticals' MGL-3196(I) selectively acts on THR-β, has obvious effect in patients with non-alcoholic steatohepatitis (NASH), and has little side effects on the liver. It is a safe lipid-lowering (LDL, LDL-C) and TG) drugs, which are currently in the key development stage of clinical trials (J.Med.Chem.2014, 57, 3912-3923). F.Hoffmam-La Roche Company introduced a substituent at the 2-position of pyridazinone to obtain a series of compounds II. Compound II not only has the effect of treating NASH, but also has the function of anti-thyroid cancer (WO2009037172A1). The substituent introduced by Sichuan Haisco Pharmaceutical Co., Ltd. on the 2-position of pyridazinone is an ether substituent, and the corresponding derivative III can be used for the treatment of obesity, hyperlipidemia, hypercholesterolemia, diabetes and other related diseases (CN110938094A ). Jiaxing Tecoluo Biotechnology Co., Ltd. invented IV, which can be used to treat male hormone hereditary alopecia or seborrheic alopecia or alopecia caused by chemotherapy (CN108727344A).

Selective agonists of THR-β represented by MGL-3196 can significantly reduce liver fat and fibrosis in NASH patients, as well as LDL cholesterol, triglycerides and lipoproteins. Therefore, selective agonists of THR-β are ideal candidates for reducing cardiovascular risk in NASH patients and in patients with moderate statin doses or dyslipidemia intolerant to statins (European Heart Journal, Volume 39, Issue suppl_1, 1August 2018, 566.P5387).

Since the physiological effects of thyroid hormones affect almost every organ system, it is still necessary to further improve the absorption, distribution, transport and metabolism of drugs in the body, as well as improve the bioavailability and the selectivity of drugs on target sites, in order to enhance the Efficacy, reduce the toxic and side effects of drugs, prolong the action time, so as to meet the needs of society and better benefit mankind.

SUMMARY OF THE INVENTION

A compound of general structural formula (I), or a pharmaceutically acceptable salt, polymorph, tautomer, stereoisomer, hydrate, solvate or any isotopic derivative thereof:

General structural formula (I)

in,

R ¹ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, -OH, ethynyl, halogen, amino, alkyl, alkenyl, haloalkyl, haloalkene radical, heteroalkyl, heterocycloalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxy, alkane Oxyalkyl, alkenyloxy, -alkynyl, -alkylalkynyl, -alkynyloxy, -alkylalkynyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfo Acyl, alkylsulfonyl, alkylsulfinyl, aminosulfonyl, acyl; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, -CF ₃ , Alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino.

R is selected from ^: H, alkyl, alkenyl, haloalkyl, haloalkenyl, heteroalkyl, heterocycloalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heterocycle Arylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxy, alkoxyalkyl, alkylamino, alkylaminocarbonyl, -alkynyl, -alkylalkynyl, -alkynyloxy, -alkane alkynyloxy, sulfonyl, alkylsulfonyl, alkylsulfinyl, aminosulfonyl; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, _-CF3 , alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino.

R ³ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, OH, -SR ⁷ , halogen, amino, alkyl, alkoxy, alkoxyalkyl, hetero Alkyl, heterocycloalkyl, arylalkyl, cycloalkyl, heterocycloarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkenyloxy, -alkynyl, -alkylalkynyl, -alkyne Oxy group, -alkylalkynyloxy group, alkylamino group, aminoalkyl group, alkylaminocarbonyl group; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, -CF3, alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino.

Each of ^R4 and ^R5 is selected from: H, halogen, alkyl, alkoxy, alkoxyalkyl.

R ⁶ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, -OH, alkyl, halogen, carboxyl, ester, amide.

X is selected from: covalent bond, -O-, -S-, -NH-, _-SO2- , -CONH- or ( _CH2 ) _q , X- is attached to _C4- , _C5 of benzimidazole ring _- , C6- and C7 _- position carbon atoms, q is 1, 2 or 3.

In certain specific embodiments, the preferred R1 is: H, halogen, ^C1 -C4 alkyl.

In another specific embodiment, the preferred R2 is: H, C1 ^- C4 alkyl.

In another specific embodiment, the preferred ^R3 is: H, halogen, C1-C6 alkyl.

In another specific embodiment, the preferred X is: -O-.

^In another specific embodiment, the preferred R4 and R5 are each Br, wherein R4 and ^R5 are attached to the _C3- and ^C5- _positions , respectively, ^on the corresponding benzene ring.

In another specific embodiment, the preferred R ⁴ and R ⁵ are each Cl, wherein R ⁴ and R ⁵ are attached to the C ₃ - and C ₅ - positions, respectively, on the corresponding benzene ring.

^In another specific embodiment, the preferred R6 is: -CN.

In addition to the compounds represented by the general structural formula (I), the present invention also includes pharmaceutically acceptable salts, polymorphs, tautomers, stereoisomers, hydrates and solvates of these compounds or isotopic derivatives, pharmaceutically acceptable prodrugs and pharmaceutically active metabolites, and pharmaceutically acceptable salts of these metabolites.

The present invention includes any pharmaceutical dosage form formed by the compound represented by the general structural formula (I) and a pharmaceutically acceptable diluent, excipient or carrier.

The present invention also relates to methods of preparing these compounds. These compounds can be used as thyroid hormone receptor beta-selective and/or tissue-selective agonists, resulting in modest reductions in body weight, lipids, cholesterol, and lipoproteins, effects on cardiovascular function or on the hypothalamic/pituitary/thyroid axis. Normal function has reduced impact. These compounds are useful in the treatment of metabolic diseases such as obesity, hyperlipidemia, hypercholesterolemia, diabetes and other conditions and diseases such as hepatic steatosis and NASH, atherosclerosis, cardiovascular disease, hypothyroidism, thyroid cancer, thyroid disease, and related conditions and diseases.

In another specific embodiment, the diseases that the compound represented by the general structural formula (I) can treat are: various lipid disorders, metabolic syndrome, obesity, atherosclerosis or diabetes.

In another specific embodiment, the diseases that the compound represented by the general structural formula (I) can treat are: NASH and Duchenne muscular dystrophy syndrome.

In another specific embodiment, the diseases that the compound represented by the general structural formula (I) can treat are: primary biliary cirrhosis and cholangitis, including senile dementia and tumors.

In another specific embodiment, the diseases that the compound represented by the general structural formula (I) can treat are: increase the energy and activation performance of T lymphocytes to enhance immune function, and can transform tumor cells into fat cells, reduce cancer transfer etc.

In another specific embodiment, the compound represented by the general structural formula (I) can treat visceral fat type obesity.

In another specific embodiment, compounds represented by general structural formula (I) can treat thyroid diseases, including thyroid cancer.

One or more embodiments of the present application also provide a method of treating and/or prophylactically treating a disease regulated by thyroid hormone, comprising administering a compound of the present application to a subject in need thereof.

One or more embodiments of the present application also provide methods of treating and/or prophylactically treating obesity, diabetes, NASH, cardiovascular disease, hypothyroidism, or thyroid cancer modulated by thyroid hormone, comprising adding the present application's The compound is administered to a subject in need.

One or more embodiments of the present application also provide methods of treating and/or prophylactically treating thyroid hormone-mediated hyperlipidemia or atherosclerosis, comprising administering a compound of the present application to a subject in need thereof.

One or more embodiments of the present application also provide methods of treating and/or prophylactically treating hypercholesterolemia mediated by thyroid hormone, comprising administering a compound of the present application to a subject in need thereof.

One or more embodiments of the present application also provide compounds for use in the treatment and/or prophylactic treatment of diseases regulated by thyroid hormones.

One or more embodiments of the present application also provide compounds for the treatment and/or prophylactic treatment of obesity, diabetes, NASH, cardiovascular disease, hypothyroidism, or thyroid cancer modulated by thyroid hormones.

One or more embodiments of the present application also provide compounds for use in the treatment and/or prophylactic treatment of thyroid hormone-mediated hyperlipidemia or atherosclerosis.

One or more embodiments of the present application also provide compounds for use in the treatment and/or prophylactic treatment of thyroid hormone-mediated hypercholesterolemia.

Description of drawings

Fig. 1 shows the test results of the blood TG (triglyceride) index in the in vivo drug effect test.

Figure 2 shows the test results of food utilization in the in vivo efficacy test.

detailed description

definition

As used herein, the term "compound of the present invention" refers to a compound of formula (I). The term also includes the various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula (I).

The term "unsubstituted" as used herein means no substituents or only hydrogen substitution.

Some terms used in the present invention are defined as follows:

"Halogen" refers to fluorine, chlorine, bromine and iodine.

"Alkyl" when taken as a group or part of a group refers to a straight or branched chain aliphatic hydrocarbon group. Preferred alkyl groups are C1-C14 alkyl groups; more preferred are: C1-C10 alkyl groups; most preferred are C1-C6 alkyl groups, unless otherwise specified. Examples of straight or branched C1-C6 alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, hexyl Wait.

"Alkylamino" includes both monoalkylamino and dialkylamino unless otherwise indicated. "Monoalkylamino" refers to: a group of (alkyl-NH)-; "dialkylamino" refers to: a group of ((alkyl)2N) _- . Wherein, alkyl is as defined herein. The alkyl group is preferably a C1-C6 alkyl group. Examples include, but are not limited to: N-methylamino, N-ethylamino, N-isopropylamino, N,N-(diethyl)amino, and the like.

"Aminoalkyl" refers to: (amino-alkyl)- group. Wherein, alkyl is as defined herein. Examples include, but are not limited to: aminoethyl, 1-aminopropyl, 1-aminopropyl, and the like.

"Arylamino" includes both mono-arylamino and di-arylamino unless otherwise specified. Mono-arylamino refers to: a group of (aryl-)NH-; di-arylamino refers to a group of formula (aryl) _2N- ; see the relevant section herein for the definition of aryl.

"Acyl" includes (alkyl-CO)- groups and (aryl-CO)- groups unless otherwise specified. Wherein alkyl or aryl are as defined herein. Examples of acyl groups include, but are not limited to: acetyl, propionyl, isobutyryl, benzoyl, and the like.

"Amido" includes (alkyl-CONH)- groups and (aryl-CONH)- groups unless otherwise specified. Wherein, the alkyl group or the aryl group refer to the relevant definitions herein. Examples of amide groups include, but are not limited to, acetamido, propionamido, butanamido, isobutyramido, benzamido, and the like.

"Alkenyl" when used as a group or part of a group refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, which may be straight or branched. Preference is given to alkenyl groups having C2-C14. C2-C12 are better; most preferred are the C2-C6 alkenyl groups. The group may contain multiple double bonds in its backbone and may each be E or Z in conformation. Examples of alkenyl groups include, but are not limited to: vinyl, propenyl, and the like.

"Alkoxy" refers to a (alkyl-O)- group. Wherein, alkyl is as defined herein. C1-C6 alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and the like.

"Alkenyloxy" refers to a (alkenyl-O)- group. where alkenyl is as defined herein. Alkenyloxy of C1-C6 is preferred.

"Alkynyloxy" refers to a (alkynyl-O)- group. Wherein alkynyl is as defined herein. C1-C6 alkynyloxy groups are preferred.

"Alkoxycarbonyl" refers to a group of (alkyl-O-C(O))-. Wherein, alkyl is as defined herein. Preferred alkyl groups are C1-C6 alkyl groups. Examples thereof include, but are not limited to: methoxycarbonyl, ethoxycarbonyl, and the like.

"Alkylsulfinyl" refers to a group of (alkyl-S(O))-. where alkyl is as defined herein. Preferred alkyl groups are C1-C6 alkyl groups. Alkylsulfinyl groups include, but are not limited to, methylsulfinyl, ethylsulfinyl, and the like.

"Alkylsulfonyl" refers to a group of (alkyl-S(O) ₂ -O)-. where alkyl is as defined herein. The preferred alkyl group is a C1-C6 alkyl group. Examples include, but are not limited to, methanesulfonyl, ethanesulfonyl, and the like.

"Alkylaminocarbonyl" refers to an alkylamino-carbonyl group. Wherein, alkylamino is as defined herein.

"Cycloalkyl" refers to a saturated or partially saturated monocyclic, fused or spirocyclic carbocyclic ring. Rings of 3-9 carbon atoms are preferred. Examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

"Cycloalkylalkyl" refers to a cycloalkyl-alkyl group. Wherein, cycloalkyl and alkyl moieties are as defined herein. Monocycloalkylalkyl groups include, but are not limited to: cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, and the like.

"Heterocycloalkyl" refers to a cycloalkyl group containing at least one heteroatom selected from N, S, O. It preferably contains 1 to 3 (1, 2 or 3) heteroatoms. Preferred rings are 3-14 membered rings, more preferred rings are 4-7 membered rings. Heterocycloalkyl includes, but is not limited to: pyrrolidinyl, dihydropyrrolyl, tetrahydropyrrolyl, dihydropyrazolyl, piperidinyl, morpholinetetrahydrofuranyl, tetrahydrothiofuranyl, tetrahydropyran Base et al.

"Heterocycloalkenyl" refers to a heterocycloalkyl group containing at least one double bond. Heterocycloalkyl is as defined herein.

"Heterocycloalkylalkyl" refers to the group of: (heterocycloalkyl-alkyl)-. Wherein, heterocycloalkyl and alkyl moieties are as defined herein. Heterocycloalkylalkyl groups include, but are not limited to: (2-tetrahydrofuranyl)methyl, (2-tetrahydrothiofuranyl)methyl, and the like.

"Heteroalkyl" means a straight or branched chain alkyl group containing at least one or more (eg 1, 2 or 3) heteroatoms selected from S, O and N in the main chain . Chains containing 2-14 atoms are preferred. Heteroalkyl groups include, but are not limited to, ethers, thioethers, alkyl esters, second or third alkylamines, alkylsulfinic acids, and the like.

"Aryl" as a group or part of a group refers to: (1) an aromatic monocyclic or fused ring; preferably an aromatic carbocyclic ring with 5-12 carbon atoms (ring atoms are all carbon atoms) ring structure). Examples of aryl groups include, but are not limited to: phenyl, naphthyl; (2) a partially saturated carbocyclic ring may be attached, for example: phenyl and C5-7 cycloalkyl or C5-7 cycloalkenyl groups are fused to each other combined to form a ring structure. Examples include, but are not limited to: tetrahydronaphthyl, indenyl, or indenyl, and the like. Aryl groups may be substituted with one or more substituents.

"Arylalkenyl" refers to the group of: (aryl-alkenyl)-. where aryl and alkenyl are as defined herein. Exemplary arylalkenyl groups include, but are not limited to: phenylpropenyl and the like.

"Aralkyl" refers to: (aryl-alkyl)- group. Wherein, the aryl and alkyl moieties are as defined herein. Exemplary aralkyl groups include, but are not limited to, benzyl, phenethyl, 1-naphthylmethyl, and the like.

"Cycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system. It contains at least one carbon-carbon double bond and preferably has 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include, but are not limited to: cyclopentene, cyclohexene, or cycloheptene. Cycloalkenyl groups may be substituted with one or more substituents.

"Heteroaryl" refers to a monocyclic or fused polycyclic aromatic heterocycle. Preference is given to 5-7 membered aromatic rings containing one or more (eg 1, 2 or 3) heteroatoms selected from N, O or/and S. Typical heteroaryl substituents include, by way of example, but not limited to: furyl, thienyl, pyrrole, pyrazole, triazole, thiazole, pyridine, pyrimidine, pyrazine, indole, benzimidazole, and the like.

"Heteroaralkyl" refers to the group of: (heteroaryl-alkyl)-. Wherein, the heteroaryl and alkyl moieties are as defined herein. Exemplary heteroaralkyl groups include, but are not limited to: 2-furanmethyl, 3-furanmethyl, 2-pyridylmethyl, and the like.

The present invention includes compounds represented by the general structural formula (I) and various possible isomeric forms thereof. Including: non-spiroisomers, mirror isomers, tautomers and geometric isomers of "E" or "Z" configuration isomers, etc. Any chemist with certain basic knowledge can isolate the above optically pure or stereoisomerically pure compounds.

The present invention includes compounds represented by the general structural formula (I) and mixtures of possible racemates or/and enantiomers and/or/and diastereomers thereof.

In addition, the compound represented by the general structural formula (I) also covers the solvated and unsolvated forms of the compound in application. Accordingly, each formula includes compounds of the indicated structures, including hydrates, solvates, and anhydrous forms thereof.

The compound represented by the general structural formula (I) of the present invention also covers various crystal forms and amorphous forms of the compound in application. Therefore, the above-mentioned various crystal forms include, but are not limited to, various hydrate crystal forms, various solvate crystal forms, and various anhydrous compound crystal forms, and the like.

As used herein, the term "pharmaceutically acceptable salt" means, within the scope of sound medical judgment, suitable for contact with human and lower animal tissues without undue toxicity, irritation, allergy, etc., and is compatible with reasonable The benefit/hazard ratio of those salts is commensurate. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. in J. Pharmaceutical Sciences (1977) 66: 1-19 describe pharmaceutically acceptable salts in detail. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric and perchloric acids, or salts formed with organic acids such as acetic, oxalic, Maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Also included are salts formed using methods conventional in the art, eg, ion exchange methods. Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, camphor acid salt, camphorsulfonate, citrate, cypionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerin Phosphate, Gluconate, Hemisulfate, Heptanoate, Caproate, Hydroiodide, 2-Hydroxy-ethanesulfonate, Lactobate, Lactate, Laurate, Lauryl Sulfate , malate, maleate, malonate, mesylate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoic acid Salt, Pectin Acetate, Persulfate, 3-Phenylpropionate, Phosphate, Picrate, Pivalate, Propionate, Stearate, Succinate, Sulfate, Tartrate, Thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Pharmaceutically acceptable salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations with counter ions such as halides, hydroxides, carboxylates, sulfates, phosphates, Nitrates, lower alkyl sulfonates and aryl sulfonates.

The term "solvate" refers to a complex in which a compound of the present invention coordinates with solvent molecules to form a complex in specified proportions. "Hydrate" refers to a complex formed by the coordination of a compound of the present invention with water.

The compounds of the present invention may contain one or more asymmetric centers, and thus may exist in various "stereoisomeric" forms, eg, enantiomeric and/or diastereomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers, or geometric isomers (eg, cis and trans isomers), or may be in the form of a mixture of stereoisomers, Include racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be separated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and chiral salt formation and crystallization; or preferred isomers can be separated by prepared by asymmetric synthesis.

The pharmacological effect can be embodied by the following but not limited to the following description.

The ligand binding domain of THR-β (amino acids 148-410) (H6-THR-β) and the ligand binding domain of THR-α (amino acids 202-461) (H6-TRα) were cloned into E. coli for expression In the vector pET28a (Novagen, Milwaukee, WI), the vector contains six His sequences at the N-terminus. The resulting recombinant Hex-His-tagged protein was produced in E. coli BL21(DE3) cells. Cells were grown in Terrific Broth (home-made Bacto tryptone (3.3%, w/v), Difico yeast extract (2.0%, w/v) and NaCl (0.5%, w/v) medium) , using a shake flask, incubated in 0.2 mM IPTG at 25°C for 24 hours, harvested or treated with 5 volumes of buffer A (0.05M Tris, 0.3M NaCl, 1% w/v betaine, 0.01M imidazole, 0.02M b-mercaptoethanol, pH 8.0) dissolved. Lysozyme (1.0 mg/ml, Sigma) and CompleteProtease Inhibitor Cocktail (Roche Diagnostics Gmbh) were added to the slurry, and the solution was sonicated for one minute, 5 times at 4°C. The suspension was centrifuged at 127,300 RCF in a Ti45 Beckman rotor for 2 hours and the supernatant was loaded onto a NI NTAAgarose (Quigen 30210) column. After washing with buffer A, H6-THR-β or H6-THR-α was eluted with buffer A containing 0.25M imidazole.

Mix 30 μl of H6-THR-β (50 nM) or H6-THR-α in 50 mM Hepes, pH 7.0, 1 mM DTT, 0.05% NP40 and 0.2 mg/ml BSA (binding buffer) with an equal volume of EE-RxRα (50 nM) in binding buffer was mixed. Then 6 microliters of T3 (0-14.8 uM) or test compound (0-1.2 mM) in DMSO were added and the solution was incubated at 37°C for 30 min. 30 microliters of biotin-GRIP peptide (Biotin-Aca-HGTSLKEKHKILHRLLQDSSSPVDL-CONH2) (100 nM) in 30 ul of binding buffer plus 5% DMSO was then added and the solution was incubated at 37°C for 30 min. 30 μl of a solution containing 12 nM europium-conjugated anti-hexa-His antibody and 160 n MAPC-conjugated streptavidin in 50 mM Tris, pH 7.4, 100 mM NaCl and 0.2 mg/ml BSA was added and the solution was Incubate overnight at 4°C. Aliquots (35ul/sample) were transferred to 384 well black microtiter plates. HTRF signals were read on a Victor5 reader (PerkinElmer Life and Analytical Sciences). EC50 was calculated.

The term "prodrug" includes a class of compounds which may themselves be biologically active or inactive and, when administered in an appropriate manner, undergo a metabolic or chemical reaction in the human body to convert into a class of compounds of formula (I), or A salt or solution of a compound of (I). The prodrugs include (but are not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino compounds, carbamates, azo compounds, phosphorus Amide, glucoside, ether, acetal and other forms.

As used herein, the term "subject" includes, but is not limited to, humans (ie, male or female of any age group, eg, pediatric subjects (eg, infants, children, adolescents) or adult subjects (eg, Young adults, middle-aged adults, or older adults)) and/or non-human animals, eg, mammals, eg, primates (eg, cynomolgus monkeys, rhesus monkeys), cows, pigs, horses , sheep, goats, rodents, cats and/or dogs. In some embodiments, the subject is a human. In other embodiments, the subject is a non-human animal.

Unless otherwise specified, the term "treatment" as used herein includes the effect that occurs when a subject suffers from a particular disease, disorder or condition, which reduces the severity of, or delays or slows down, the disease, disorder or condition or development of a disorder ("therapeutic treatment"), and also includes effects that occur before a subject begins to suffer from a particular disease, disorder, or disease ("prophylactic treatment").

"Combination" and related terms refer to the simultaneous or sequential administration of the therapeutic agents of the present invention. For example, a compound of the present invention may be administered concurrently or sequentially with another therapeutic agent in separate unit dosage forms, or concurrently with another therapeutic agent in a single unit dosage form.

"Pharmaceutically acceptable excipient" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated together. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin. ), buffer substances such as phosphate), glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, chloride Sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycols and lanolin.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include, but are not limited to: oral, duodenal, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical Medicine.

Solid dosage forms for oral administration include capsules, tablets, pills and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or solubilizers, for example, starch , lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as, for example, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, for example, paraffin; (f) ) absorption accelerators, eg, quaternary amine compounds; (g) wetting agents, eg, cetyl alcohol and glyceryl monostearate; (h) adsorbents, eg, kaolin; and (i) lubricants, eg, talc , calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions may be in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric substances and waxes. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert release agents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

The present invention provides a method of treating a disease-related disorder caused by THR-beta in a subject in need thereof, comprising the steps of: administering to the subject in need thereof a compound of the present invention, or a pharmaceutically acceptable salt thereof, Stereoisomers, solvates, hydrates, crystal forms, prodrugs or isotopic derivatives, or administer the pharmaceutical compositions of the present invention.

The compounds of the present invention can be used to treat diseases caused by THR-β: any acute or chronic liver disease involving pathological destruction, inflammation, degeneration and/or proliferation of liver cells, various lipid disorders, metabolic syndrome.

The present invention also relates to methods of preparing these compounds, which can be used as THR-beta-selective and/or tissue-selective agonists, resulting in appropriate reductions in body weight, lipids, cholesterol, and lipoproteins, effects on cardiovascular function or The normal function of the hypothalamic/pituitary/thyroid axis has reduced effects. These compounds are useful in the treatment of metabolic diseases such as obesity, hyperlipidemia, hypercholesterolemia, diabetes and other conditions and diseases such as hepatic steatosis and NASH, atherosclerosis, cardiovascular disease, hypothyroidism, thyroid cancer, thyroid Diseases, Diabetic NASH, Duchenne Muscular Dystrophy, and related conditions and diseases.

Preparation of Nitrile Triazine Derivatives

The compounds represented by the general structural formula (I) can be synthesized using the synthetic routes and synthetic methods discussed below. The raw materials used are convenient and easy to obtain. However, the synthetic route and synthetic method used in the present invention can be widely used in the synthesis of analogs, and only the starting materials need to be changed. For example, in the synthesis of the compounds not described in detail in the examples herein, as long as the starting materials are replaced with the starting materials of the corresponding target compounds, according to chemical common sense, the desired target compounds can be synthesized by slightly changing the reaction conditions when necessary.

The synthetic routes used in the present invention may also be adapted to prepare intermediates or target compounds other than those in the specific embodiments. The reagents or intermediates used may or may not be protected with protective groups. A list of suitable protecting groups in organic synthesis can be found in G.M. Peter's Protective Groups in Organic Synthesis, Wiley, 2007. Alternatively, other reactions disclosed herein or known in the art.

Reagents that can be used to synthesize target compounds can be obtained or prepared according to known techniques.

In the following examples, all temperatures are in degrees Celsius unless otherwise indicated.

Various starting materials and reagents were obtained commercially. Suppliers include but are not limited to: Aldrich Chemical Company, Lancaster Synthesis Ltd, etc. Commercially available starting materials and reagents were used without further purification unless otherwise indicated.

Glassware is oven dried and/or heat dried. Reactions were followed on glass silica gel-60F254 plates (0.25 mm) (TLC). Analytical thin layer chromatography and developed with appropriate solvent ratios (v/v). The end point of the reaction was the consumption of starting material on TLC (J. Org. Chem., Vol. 43, No. 14, 19782923-2925).

Typically, the follow-up is to double the volume of the reaction solution with the solvent used for the reaction, and then perform three extractions with 25% of the total volume of extraction solvent, unless otherwise specified. The product-containing extract was dehydrated over anhydrous sodium sulfate, filtered on a rotary evaporator, and the solvent was evaporated under reduced pressure, taking care to remove the solvent in vacuo. Finally, the target compound was obtained by flash column chromatography.

¹ H NMR spectra were obtained with a Bruker instrument (200-400 MHz) and chemical shifts are expressed in ppm. Chloroform was used as reference standard (7.25 ppm) or tetramethylsilane internal standard (0.00 ppm). Other solvents commonly used for NMR can also be used, if desired. ¹ H NMR representation: s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, dd=doublet of doublet, dt=doublet of triplet. When coupling constants are provided, they are in Hz.

Mass spectrum is obtained by LC/MS instrument, and the ionization mode can be ESI or APCI. All melting points are uncorrected.

The following examples are merely intended to illustrate the synthesis of the specific compounds of the invention. But there are no restrictions on the synthesis method. Compounds not listed below can also be prepared by using the same synthetic route and synthetic method as below, selecting appropriate starting materials, and adjusting reaction conditions with appropriate common sense where necessary.

synthesis

The target compound in the general structural formula (I) can be synthesized by the method shown in Synthesis Scheme_1. The VII obtained after the amination and reduction of the substituted o-fluoronitrobenzene derivative V. Cyclization of intermediate VII with a suitable acid or aldehyde gives VIII. The corresponding alcohol or sulfhydryl derivative IX obtained by removing the methyl group from the latter is reacted with the fluoro derivative to obtain the key intermediate XI. After reduction, diazotization and cyclization of XI, the target compound represented by the general structural formula (I) can be obtained.

Synthetic route_1

In the general structural formula (I), when R ¹ , R ³ =H, and R ² =methyl, the corresponding compound clearly shows that compound XXIII can be synthesized by the method shown in Synthetic Scheme_2.

Synthetic route_2

Specifically, when R ¹ , R ² =H, R ³ =methyl, and X=O, S, the compound XXIII shown in the general structural formula (I) can use a suitable o-fluoronitrobenzene derivative XV is a raw material, and under the action of triethylamine, compound XVI is obtained. Reduction in the presence of a suitable catalyst such as palladium carbonyl affords XVII. Then, intermediate XVII is heated with formic acid to cyclize to give XVIII. Under the action of boron tribromide, XVIII is demethylated to obtain the corresponding alcohol or mercapto derivative XIX. The latter is condensed basicly with the appropriate compound X to give the corresponding XX. XX undergoes reduction (for example, reduction under iron powder), diazotization, and then reacts with XIII to obtain XXII. Under the action of acetic acid-sodium acetate, the target compound XXIII is obtained by cyclization.

Synthetic route_3

In the general structural formula (I), when R1, R2 ⁼ H, ^R3 =methyl, and X= ^CH2 , the corresponding target _compound XXV can be synthesized by the method shown in synthetic route_3. Specifically, the compound shown in the general structural formula (I) can use a suitable o-fluoronitrotoluene derivative XVI as a raw material, and under the action of triethylamine, compound XVII can be obtained. Reduction under the catalysis of a suitable catalyst (eg palladium carbonyl) gives XVIII. Then, intermediate XVIII is heated with formic acid to cyclize to give the corresponding XIX. Under the action of NBS, XIX was converted to the bromo-substituted derivative XX. The latter is condensed with a suitable boronic acid derivative XXI to give XXII. XXII undergoes reduction (for example, reduction under iron powder), diazotization, and then reacts with XIII to obtain XXIV. Under the action of acetic acid-sodium acetate, the target compound XXV is obtained by cyclization.

The content of the present invention is further illustrated below in conjunction with examples. The purpose is to make those skilled in the art more clearly understand and practice the specific content of the present invention. However, the scope of protection of the present invention is not limited only to these examples.

Example 1

2-(3,5-Dibromo-4-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)phenyl)-3,5-dioxo-2,3 , Preparation of 4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (1).

The compound of Example 1 (Compound 1) can be carried out by the method of the above-mentioned synthetic route_1. Specifically, as long as the desired raw material is selected, it can be synthesized by a specific route represented by the following formula.

The synthesis of step-1 5-methoxybenzene-N-methyl-2-nitroaniline (VI-1)

In a sealed tube, V-1 (5.13 g, 30.0 mmol), tetrahydrofuran (5 mL), triethylamine (9.10 g, 90.0 mmol) and 2M methylamine in furan (30 mL, 60 mmol) (40.0 g, 0.2 M) were added sequentially mol). Heat at 60°C overnight. It was then cooled to room temperature and the solvent was removed under reduced pressure. 100 mL of ethyl acetate was added, washed twice with water (50 mL) each time; washed with brine once (50 mL), and then dried over Na ₂ SO ₄ . After removing the solvent under reduced pressure, it was recrystallized from petroleum ether-ethyl acetate (petroleum ether/ethyl acetate, 1/1, 15 mL) to obtain the title compound VI-1 (white solid; 3.95 g; yield: 72%) . ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.28 (br s, 1H), 8.13 (d, J=9.2 Hz, 1H), 6.23 (dd, J=9.6, 2.4 Hz, 1H), 6.12 (d, J=2.4Hz, 1H), 3.88 (m, 3H), 3.00 (d, J=5.2Hz, 3H).

Step-2 Synthesis of 5-methoxy-N ¹ -methyl-1,2-phenylenediamine (VII-1)

To a solution of VI-1 (3.95 g, 21.7 mmol) in methanol (50 mL) was added Pd/C (10%, 800 mg). The reaction was bubbled with hydrogen (1 atm) with stirring and heated at 50°C for 6 hours. The reaction was cooled to room temperature, filtered, and concentrated to give VII-1 (brown solid; 3.25 g; yield: 98%). ¹ H NMR (400 MHz, CDCl ₃ ): δ 6.34 (d, J=8.0 Hz, 1H), 6.25 (d, J=2.4 Hz, 1H), 6.18 (dd, J=8.4, 2.8 Hz, 1H), 3.76 (s, 3H), 2.84 (s, 3H).

Step-3 Synthesis of 6-methoxy-1-methyl-1H-benzo[d]imidazole (VIII-1)

VII-1 (3.25 g, 21.4 mmol) was dissolved in formic acid solution (20 mL) and heated to 50°C overnight. Cool and concentrate under reduced pressure. Neutralize with excess saturated sodium bicarbonate solution and stir for 10 minutes. Wash twice with dichloromethane-methanol (20/1, 200 mL). The washings were combined, then washed with oxygen-saturated brine, dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated to obtain an oily substance, the title compound VIII-1 (3.4 g, yield: 94%). ¹ H NMR (400 MHz, CDCl ₃ ): δ 7.75 (s, 1H), 7.67 (d, J=8.8 Hz, 1H), 6.62 (dd, J=8.4, 2.4 Hz, 1H), 6.82 (d, J = 2.8 Hz, 1H), 3.88 (s, 3H), 3.78 (s, 3H).

Step-4 Synthesis of 1-methyl-1H-benzo[d]imidazol-6-ol (IX-1)

Under nitrogen flow at -78°C, VIII-1 (3.24 g, 20.0 mmol) was added to a reaction flask containing boron tribromide in dichloromethane solution (1 M, 80 mL, 80 mmol). Then, the temperature was naturally raised to room temperature, and the reaction was carried out overnight with stirring. Concentrate and neutralize to pH=8 with saturated sodium bicarbonate solution. Extract three times with dichloromethane-methanol solution (10/1, 200 mL). The extract was washed with water and dried over Na ₂ SO ₄ . Filtration and concentration gave the title compound IX-1 (brown solid, 2.0 g, yield: 68%). ¹ H NMR (400 MHz, CDCl ₃ ): δ 9.29 (s, 1H), 7.94 (s, 1H), 7.41 (d, J=8.4 Hz, 1H), 6.82 (d, J=2.0 Hz, 1H), 6.71 (q, J=2.4 Hz, 1H), 3.72 (s, 3H).

Step-5 Synthesis of 6-(2,6-dibromo-4-nitrophenoxy)-1-methyl-1H-benzo[d]imidazole (XI-1)

In the reaction flask, IX-1 (430 mg, 2.90 mmol), dimethyl sulfoxide (10 mL), sodium carbonate (802 mg, 5.81 mmol) and X-1 (890 mg, 3.0 mmol) were sequentially added. The reaction was stirred at 50°C for 2 hours. Cool to room temperature and add ethyl acetate (100 mL). The organic phase was washed three times with 30 mL each. Washed with saturated brine (30 mL), dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated and crystallized from petroleum ether-ethyl acetate (1/1, 15 mL) to obtain the title compound XI-1 (white solid, 1.1 g, yield: 89%). ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.66 (s, 2H), 8.13 (s, 1H), 7.61 (d, J=9.2 Hz, 1H), 7.05 (d, J=2.4 Hz, 1H) ), 6.80 (dd, J=9.2, 2.8 Hz, 1H), 3.75 (s, 3H).

Step-6 Synthesis of 3,5-dibromo-4-(1-methyl-1H-benzo[d]imidazol-6-yl)oxy-aniline (XII-1)

In the reaction flask, XI-1 (1.00 g, 2.34 mmol), ethanol (50 mL), and iron powder (1.31 g, 23.4 mmol) were added successively. Under stirring, the mixture was heated to 75°C. Concentrated hydrochloric acid (2 mL) was then added dropwise. The reaction was continued at 75°C for 30 minutes. Cool to room temperature and filter. Concentrate and neutralize with saturated aqueous sodium carbonate (50 mL). Dichloromethane-methanol solution (10/1, 150 mL) was extracted three times. The extract was washed with water and dried over Na ₂ SO ₄ . Filtration and concentration gave the title compound XII-1 (white solid, 830 mg, yield: 89%). ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.08 (s, 1H), 7.55 (d, J=8.4 Hz, 1H), 6.92 (s, 2H), 6.86 (d, J=2.8 Hz, 1H), 6.69 (dd, J=8.8, 2.4Hz, 1H), 5.61 (br s, 2H), 3.73 (s, 3H).

Step_7 (E)-Ethyl-(2-cyano-2-(2-(3,5-dibromo-4-((1-methyl-1H-benzo[d]imidazol-6-yl) ) Oxygen) Benzene) Hydrazine) Acetyl Carbamate (XIV-1) Synthesis

To an aqueous solution (7.5 mL) containing XII-1 (300 mg, 0.76 mmol) was added concentrated hydrochloric acid (0.15 mL). Stir at room temperature to give a clear solution. Cool to 0°C. A solution of NaNO2 (62 _mg , 0.76 mmol) in water (1.2 mL) was added dropwise with stirring, maintaining the temperature of the reaction suspension below 5°C. After the dropwise addition was completed, stirring was continued at 0°C for 1 hour. The reaction was added dropwise to XIII-1 (142 mg, 0.910 mmol) in pyridine-aqueous solution (8 mL-3.6 mL), maintaining the reaction temperature not to exceed 5 °C. The resulting reaction solution was further stirred at 5°C for 1 hour. The reaction solution was poured into saturated sodium bicarbonate solution (20 mL), and extracted three times with dichloromethane-methanol solution (10/1, 100 mL). The extract was washed with water and dried over Na ₂ SO ₄ . Filter and concentrate. The title compound XIV-1 (white solid, (240 mg, stringency: 56%) was obtained after C18 column separation (development system: 5-95% CH ₃ CN aqueous solution, 25 min). ¹ H NMR (400 MHz, DMSO-d ₆ ) : δ10.76(s, 1H), 8.18(s, 1H), 8.16(s, 2H), 7.60(d, J=8.8Hz, 1H), 6.95(d, J=2.4Hz, 1H), 6.76( dd, J=8.8, 2.8 Hz, 1H), 4.19 (q, J=6.8 Hz, 2H), 3.75 (s, 3H), 1.27 (t, J=6.8 Hz, 3H).

Step-8 2-(3,5-Dibromo-4-((1-methyl-1H-benzo[d]imidazol-6-yl)oxy)-phenyl)-3,5-dioxo - Preparation of 2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (1).

Sodium acetate (179 mg, 2.19 mmol) was added to a suspension containing XIV-1 (310 mg, 0.550 mmol) in acetic acid (10 mL), and the resulting reaction suspension was stirred at 120° C. for 2 hours. The reaction was cooled to room temperature and concentrated. The title compound XIV-1 (white solid, 100 mg, yield: 35%) was obtained after C18 column separation (development system: 5-95% CH ₃ CN aqueous solution, 25 min). ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.16 (s, 1H), 7.97 (s, 2H), 7.61 (d, J=8.8 Hz, 1H), 7.05 (d, J=2.4 Hz, 1H) ), 6.75 (dd, J=8.8, 2.4 Hz, 1H), 3.77 (s, 3H). LC-MS [mobile phase: within 6.5 min from 80% water (0.02% _NH4OAc ) and 20% _CH3 CN to 30% water (0.02% _NH4OAc ) and 70% _CH3CN ], Rt=2.732 min; Purity: 96.19% (254 nm), MS calculated: 515.9; MS found: 516.9 [M+H] ⁺ .

Example 2-12

According to the method of Example 1, various derivatives can be synthesized as long as appropriate starting materials are changed. Examples 2-12 are some representative examples thereof (see Table 1).

Table 1

In addition, synthetic route_1 or synthetic route_2 can be selected. Referring to the method of Example 1, as long as the starting materials are appropriately selected, a wider variety of derivatives can also be synthesized, such as the compounds listed in Table 2. Just a few examples.

Table 2

Pharmacological test

I. The THR-α/β activating effect of the test compounds (Example Compounds 1, 2, 4, 5, 6, 7, 8, 9) was determined by the following method.

Test method 1

THR/RXR/GRIP1 detection

Insertion of genes expressing the ligand binding domain (amino acids 148-410) of THR-β (H6-THR-β) and the ligand binding domain (amino acids 202-461) of THR-α (H6-THR-α) into the large intestine In the Bacillus expression vector pET28a, the expression vector contains an N-terminal six-group His sequence. It was introduced into Escherichia coli BL21(DE3) strain to produce a His-tagged recombinant protein. The strains were inoculated into TB medium (medium formulation: trypsin 3.3%, yeast extract 2.%, NaCl 0.5%), IPTG with a final concentration of 0.2 mM was added, and cultured in shake flasks at 25°C 24h, after culturing, collect the bacteria in the culture, and use 5 times the volume of lysate (0.05M Tris, 0.3M NaCl, 1% W/V betaine, 0.01M imidazole, 0.02M β-mercaptoethanol, pH 8.0) to remove the collected bacteria Resuspend, add lysozyme and protease inhibitors at the same time, and perform sonication (5 times for 1 minute) under ice bath conditions (4°C). The lysed suspension was centrifuged at 127,300 RCF for 2 hours, and the supernatant was loaded onto a NI_NTA agarose (Quigen 30210) column, washed with buffer A, and eluted with buffer A containing 0.25M imidazole to elute H6-TRβ or H6- TRα protein.

The human retinol X receptor (amino acids 225-462) (RxRα) was engineered with His6 and EE (EFMPME) tags at the N-terminus, cloned into the pACYC vector, and produced His6EEtagged in E. coli protein. Add the final concentration of 0.1mM IPTG to the medium for induction, and incubate at 18°C for 18h in a shaker flask. After the culture, the bacteria in the culture were collected and lysed with 5 times the volume of lysate (0.05M Tris, 0.3M NaCl, 1 % w/v betaine, 0.01M imidazole, 0.02M β-mercaptoethanol, pH 8.0) The collected bacteria were resuspended with simultaneous addition of lysozyme and protease inhibitors, stirred at 4°C for 30 minutes, and then the suspension was incubated at 4°C Under sonication for 30s, 5 times. The suspension was collected and centrifuged at 12000 RCF for 20 min. The supernatant was collected and filtered using a 0.45 μm pore size membrane, followed by the addition of 0.5% NP-40. The His6-tagged protein was bound to NiNTA metal affinity resin and eluted, the protein concentrated and dialyzed. The His6 tag was removed from the EE-RxRα protein by thrombin digestion (10 units of thrombin per mg of protein, incubated at 25°C for 2 h). Thrombin was again removed in batches using anisole sepharose 6B, protein was concentrated and dialyzed. This protein is used in coactivating peptide recruitment assays.

THR-β/RXR/GRIP1 co-activating peptide recruitment assay

50 mM Hepes, pH 7.0, 1 mM DTT, 0.05% NP40 and 0.2 mg/mL BSA (binding buffer) were formulated into 30 μL of H6-THR-β (50 nM), then mixed with an equal volume of EE-RxRα (50 nM) buffer mix. Then 6 [mu]L of T3 (0-14.8 [mu]M) or test compound (0-1.2 mM) was added to DMSO and the solution was incubated at 37[deg.]C for 30 minutes. 30 μL of binding buffer and 30 μL of biotin-GRIP1 peptide (100 nM) in 5% DMSO were then added, and the solution was incubated at 37° C. for 30 minutes. Add a solution containing 12 nM conjugate-conjugated anti-hexahistidine His antibody and 160 nM APC-conjugated streptavidin, 30 μL (50 mM Tris, pH 7.4, 100 mM NaCl and 0.2 mg/mL BSA), 4 Incubate overnight at °C. Aliquots (35 μL/sample) were transferred to 384-well black microtiter plates. HTRF signals were read on a Victor 5 reader (PerkinElmer Life and Analytical Sciences).

THR-α/RXR/GRIP1 co-activating peptide recruitment assay

This assay is essentially the same as the THR-β/RXR/GRIP1 coactivating peptide recovery technique described above, except that 125 nM 6-THR-α, 125 nMMEE-RxRα and 250 nM biotin-GRIP1 are used.

Test method 2

Methods of testing compounds for THR-α/β activating activity at the cellular level in vitro:

Compounds were transferred to a 384-well plate using an ECHO liquid workstation, with 10 gradient concentrations of each compound, 3-fold dilution, and triplicate wells. Plate 1.5 x ¹⁰⁴ DMEM cultured cells (TR beta-UAS-bla HEK 293T Cell) or 1.0 x 104 TR alpha ^- UAS-bla HEK 293T Cell in a 384-well plate. HEK 293T-TR beta was incubated in the incubator for 16 hours, HEK 293T-TR alpha was incubated in the incubator for 24 hours, and LiveBLAzer ^™ -FRET B/G(CCF4-AM) substrate was added to the cell plate for detection of intracellular beta -Lactamase expression, the product will generate fluorescence at 447nm under excitation at 409nm, if β-lactamase is not expressed, under excitation at 409nm, it will directly generate fluorescence at 520nm by FRET, by detecting the ratio of the two fluorescence (blue /green, 460nm/530nm) to determine the binding of the compound to the protein, so as to calculate the EC50 of the compound. In each experiment, the reference compounds triiodothyronine (T3) and MGL3196 will be used as positive controls for the experiment. The calculation of the Z factor (greater than 0.5) will be used to monitor the stability of each experiment.

II. Test Results

Using the aforementioned method, some test results are summarized in Table 3.

table 3

In vivo efficacy test

I. Test method:

In this experiment, golden hamsters (80-110g, 5-6 weeks old, male, Beijing Weitonglihua, license number SCXK (Jing) 2016-0011) were fed with high-fat diet to produce a hyperlipidemia model, and then compound 1 was administered orally. and MGL3196 positive control drug, the content of TG (triglyceride) in plasma was determined by automatic biochemical analyzer, and the efficacy of compound 1 in the treatment of non-alcoholic fatty liver (NASH) was evaluated according to the determination results.

After the model was successfully established, the experimental animals were divided into 5 groups according to blood lipid levels, namely:

Group 1: blank control group, 6 animals, common feed, given vehicle at the same time, 10 mL/kg.

Group 2: model control group, 8 animals, fed with high-fat diet, and given vehicle at the same time, 10 mL/kg.

Group 3: low-dose group, 8 animals, high-fat feed, 5 mg/kg, 10 mL/kg.

Group 4: high-dose group, 8 animals, high-fat feed, 15 mg/kg, 10 mL/kg.

Group 5: positive control group (MGL3196), 8 animals, high-fat diet, dose of 15 mg/kg, 10 mL/kg.

Mode of administration: oral administration, once a day, time interval 24h, continuous administration for 4 weeks, during the experiment, the animals were weighed twice a week, and the remaining amount of feed and added amount were recorded before each blood test, which was used to calculate The food intake of animals in each group during the administration period.

II. Test results:

1. Blood TG (triglyceride) index: see Figure 1;

2. Food utilization: see Figure 2.

Food utilization represents the weight gain in grams per 100g of feed, food utilization = weight gain (g)

/Food intake (100 g) x 100%. Compound 1 significantly reduced blood glycerol compared with the model group during 4 weeks of administration

Triester levels.

For food availability, compound 1 (Cpd1) had a significant decrease and MGL3196 had a certain increase. It shows that compound 1 can reduce the conversion of fat in food into body weight by increasing energy consumption under the condition of ingesting the same food, especially fat, which just confirms the mechanism of action of compound 1 as a TRβ agonist.

The details described in the specific examples of the invention are not intended to be inferred to be limiting. Various synonyms and modifications may be made without departing from the spirit and scope of the invention, and the synonymous embodiments are known to be a part of this invention.

Claims (19)

1. A compound having the structure of general formula (I), or a pharmaceutically acceptable salt, polymorph, tautomer, stereoisomer, hydrate, solvate or isotopic derivative thereof:

   

   General structural formula (I)

   in,

   R ¹ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, -OH, ethynyl, halogen, amino, alkyl, alkenyl, haloalkyl, haloalkene radical, heteroalkyl, heterocycloalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heteroarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxy, alkane Oxyalkyl, alkenyloxy, -alkynyl, -alkylalkynyl, -alkynyloxy, -alkylalkynyloxy, alkynyloxy, amino, alkylamino, aminoalkyl, alkylaminocarbonyl, sulfo Acyl, alkylsulfonyl, alkylsulfinyl, aminosulfonyl, acyl; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, -CF ₃ , Alkyl, alkenyl, alkynyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino;

   R is selected from ^: H, alkyl, alkenyl, haloalkyl, haloalkenyl, heteroalkyl, heterocycloalkyl, arylalkyl, cycloalkyl, aryl, heteroaryl, heterocycle Arylalkyl, heterocycloalkyl, heterocycloalkenyl, alkoxy, alkoxyalkyl, alkylamino, alkylaminocarbonyl, -alkynyl, -alkylalkynyl, -alkynyloxy, -alkane alkynyloxy, sulfonyl, alkylsulfonyl, alkylsulfinyl, aminosulfonyl; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, -CF ₃ , alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino;

   R ³ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, OH, -SR ⁷ , halogen, amino, alkyl, alkoxy, alkoxyalkyl, hetero Alkyl, heterocycloalkyl, arylalkyl, cycloalkyl, heterocycloarylalkyl, heterocycloalkyl, heterocycloalkenyl, alkenyloxy, -alkynyl, -alkylalkynyl, -alkyne Oxy group, -alkylalkynyloxy group, alkylamino group, aminoalkyl group, alkylaminocarbonyl group; in the above groups, each can be unsubstituted or substituted by one or more substituents, these substituents include: halogen, -CF ₃ , alkyl, alkenyl, alkynyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, amino, alkylamino, cycloalkylamino;

   R ⁴ and R ⁵ are each selected from: H, halogen, alkyl, alkoxy, alkoxyalkyl;

   R ⁶ is selected from: H, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -CO ₂ H, -OH, alkyl, halogen, carboxyl, ester, amide;

   X is selected from: covalent bond, -O-, -S-, -NH-, _-SO2- , -CONH- or -(CH2)q-, X- _C4- , C attached to the benzimidazole ring _On carbon atoms in the _5- , C6- and C7 _- positions, q is 0, 1, 2, 3.
2. ^2. A compound according to claim ¹ wherein R1, R2 and ^R3 are each H.
3. A compound according to claim ¹ , wherein R2 is methyl.
4. A compound according to claim ¹ , wherein R4 and R5 are each Br, and R4 and R5 are attached to the _C3- and ^{C5 -} _positions , respectively, ^on the corresponding benzene ring.
5. A compound according to claim ¹ , wherein R4 and R5 are each ^Cl , and R4 and R5 are attached to the _C3- and ^{C5 -} positions, respectively, _on the corresponding benzene ring.
6. A compound according to claim ¹ , wherein R4 and R5 are each F, and R4 and R5 are attached to the _C3- and ^{C5 -} _positions , respectively, ^on the corresponding benzene ring.
7. A compound according to claim ¹ , wherein R4 and R5 are each I, and R4 and _R5 are attached to the ^C2'- and ^{C6' -} positions, respectively, _on the corresponding benzene ring.
8. A compound according to claim ¹ , wherein R4 and R5 are each methyl, and R4 and R5 are attached to the _C3- and ^{C5 -} _positions , respectively, ^on the corresponding benzene ring.
9. A compound according to claim ¹ , wherein R4 and R5 are each difluoromethyl, and R4 and R5 are attached to the _C3- and ^{C5 -} _positions , respectively, ^on the corresponding benzene ring.
10. A compound according to claim ¹ , wherein R4 and R5 are each trifluoromethyl, and R4 and R5 are attached to the _C3- and ^{C5 -} _positions , respectively, ^on the corresponding benzene ring.
11. 4. A compound according to claim ¹ , wherein R6 is H.
12. A compound according to claim 1, wherein R6 is ^CN .
13. A compound according to claim ¹ , wherein R6 is lower alkyl containing 1 to 3 C atoms.
14. The compound of claim 1, wherein the compound is selected from the group consisting of the following compounds, and pharmaceutically acceptable salts thereof:

    

    

    
15. A compound according to any one of claims 1 to 14, or a combination drug with other drugs, for the treatment and/or prophylactic treatment of diseases regulated by thyroid hormones.
16. A compound according to any one of claims 1 to 14, or a combination medicament with other medicaments, for the treatment and/or prophylactic treatment of metabolic diseases regulated by thyroid hormones.
17. A compound according to any one of claims 1 to 14, or a combination drug with other drugs, for the treatment and/or prophylactic treatment of obesity, diabetes, NASH (nonalcoholic steatohepatitis, non-alcoholic fat) regulated by thyroid hormones liver), cardiovascular disease, hypothyroidism, or thyroid cancer.
18. A compound according to any one of claims 1 to 14, or a combination medicament with other medicaments, for the treatment and/or prophylactic treatment of hyperlipidemia or atherosclerosis mediated by thyroid hormones.
19. Use of a compound according to any one of claims 1 to 14, or a combination medicament with other medicaments, for the treatment and/or prophylactic treatment of hypercholesterolemia regulated by thyroid hormones.

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